New 3-phenylpropionic acid derivatives

ABSTRACT

The invention relates to new compounds, being 3-phenylpropionic acid derivatives of formula I wherein W represents COOH group or its bioisosters, or —COO—C 1 -C 4 -alkyl group; Y represents NH, N—C 1 -C 10 -alkyl, O, or S; Z represents NH, N—C 1 -C 10 -alkyl, N-aryl, N-heteroaryl, S, or O; X represents O, S, NH, N—C 1 -C 10 -alkyl, N-aryl, NSO 2 —C 1 -C 10 -alkyl, N—SO 2 -aryl, or N—SO 2 -heteroaryl; R 1  to R 8  each independently represent hydrogen atom or a substituent defined in the description; A is as defined in the description; n represents an integer from 0 to 4, inclusive; and pharmaceutically acceptable salts thereof. The compounds are the ligands of PPAR-gamma receptor and are useful as medicaments.

This application claims the benefit of Polish Patent Application No.P-371841, filed Dec. 20, 2004, which is herein incorporated by referencein its entirety.

FILED OF THE INVENTION

The present invention relates to new compounds, being 3-phenylpropionicacid derivatives, pharmaceutical compositions comprising the same, andtheir use for the treatment and/or prevention of peroxysomeproliferator-activated receptor gamma (PPARγ) mediated diseases andconditions. The compounds show the ability to bind to PPARγ receptor andmodify its activity.

THE STATE OF THE ART

More than 20 years ago, the thiazolidinedione group of compounds wasdiscovered, showing the activity in rodent models of type 2 diabetes andinsulin resistance. Although their mechanism of action was not known,the compounds have been successfully used in therapy of type 2 diabetes.Publications demonstrating that they exerted their effect via thenuclear PPAR gamma receptor were published only in the middle of 90's.Now, it is well known that intracellular receptor proteins of the PPARfamily control the expression of genes involved in the regulation oflipid-carbohydrate metabolism.

Diseases such as hyperlipidemia, atherosclerosis, obesity, and type 2diabetes become the serious concern not only for developed industrialsocieties. It is estimated that more than 150 million people worldwidesuffer from type 2 diabetes, and this number is expected to double by2025. In Poland, currently about 2 million people suffer from thisdisease, and the same number is at risk of developing it. Costs ofmedical care in diabetic patients reach 6 to 8 percent of total medicalcare budgets. At the initial stage, diabetes can be symptomless, and maybegin at any age; however, most often occurs at middle age and inelderly persons. The progress of type 2 diabetes is a result ofoverlapping of physiological disorders such as: tissue insulinresistance, insufficient pancreatic insulin production, elevated insulinproduction following intensified gluconeogenesis. Most often diabeticcomplications are microvascular changes in the retina, kidneys andnervous system, what leads to increased risk of blindness, renalinsufficiency and neuropathy. Diabetes is also the main causative factorof heart infarct and brain stroke.

PPARγ receptors, belonging to the family of nuclear receptors, play therole in the regulation of lipid metabolism and storage. They areexpressed in adipose tissue and large intestine, and are involved in thelipogenesis process. Ligands activating PPARγ receptor can enhanceinsulin effect and lower the plasma glucose level. They can be alsouseful in the management and therapy of lipid metabolism and energybalance disorders.

There are known compounds being L-tyrosine derivatives or analogues,which exert their action via modulation of the PPARγ receptor response,thus acting on the glucose metabolism, lipid hemostasis and energybalance.

In the international patent applications Nos. WO03/011834 andWO03/011814 there are disclosed N-(2-benzoylphenyl)-L-tyrosinederivatives, which have a partial PPARγ agonist activity and may beuseful in the treatment and prophylaxis of inter alia impaired insulintolerance, type 1 and 2 diabetes, dyslipidemia, disorders associatedwith syndrome X, such as hypertension, obesity, insulin resistance,hyperglycemia, atherosclerosis, myocardial ischemia, coronary heartdisease, renal diseases, as well as for improving cognitive functionsand for treating diabetic complications. The disclosed compoundsrepresent L-tyrosine derivatives wherein tyrosine hydroxyl group issubstituted with vinyl group and nitrogen in tyrosine amino group issubstituted with 2-benzoylphenyl group.

In the international patent application No. WO01/17994 there aredisclosed oxazole compounds as PPARγ antagonists, which may be useful inthe treatment of diabetes, obesity, metabolic syndrome, impaired insulintolerance, syndrome X and cardiovascular diseases, includingdyslipidemia. The compounds represent L-tyrosine derivatives whereintyrosine carboxyl group is substituted with a 5-membered heterocyclicgroup, tyrosine hydroxyl group is substituted with(5-methyl-2-phenyloxazol-4-yl)ethyl group, and nitrogen in tyrosineamino group is substituted with 2-benzoylphenyl group.

In the international patent application No. WO97/31907 there aredisclosed 4-hydroxyphenylalcanoic acid derivatives with agonisticactivity to PPARγ. Among others, there are disclosed L-tyrosinederivatives wherein tyrosine hydroxyl group is substituted with a5-membered heterocyclic group, which itself can be substituted, andnitrogen in tyrosine amino group is substituted with 2-substitutedphenyl group, including 2-benzoylphenyl group.

In the art still exists a need for new compounds-ligands of PPARγ, whichmay be useful in the treatment and/or prophylaxis of diabetes andcomplications resulting from or associated with diabetes, especiallylipid metabolism disorders and cardiovascular diseases.

THE SUMMARY OF THE IN INVENTION

The present invention relates to new compounds, 3-phenylpropionic acidderivatives of formula (I)

wherein:

-   -   W represents COOH group or its bioisosters, or —COO—C₁-C₄-alkyl        group;    -   Y represents NH, N—C₁-C₁₀-alkyl, O, or S;    -   Z represents NH, N—C₁-C₁₀-alkyl, N-aryl, N-heteroaryl, S, or O,    -   X represents O, S, NH, N—C₁-C₁₀-alkyl, N-aryl,        NSO₂—C₁-C₁₀-alkyl, N—SO₂-aryl, or N—SO₂-heteroaryl;    -   R₁, R₂, R₃, R₄, R₅, R₆, R₇ and R₈ each independently represents        hydrogen atom or a substituent selected from the group        consisiting of:    -   C₁-C₄-alkyl, C₁-C₄-alkoxy, C₃-C₇-cycloalkyl, C₃-C₇-cycloalkoxy,        C₁-C₄-thioalkoxy, C₃-C₇-cyclothioalkoxy, halogen atom,        halogen-substituted C₃-C₇-cycloalkyl, aryl, heteroaryl, —NO₂,        —CN, —SO₂—NH₂, —SO₂—NH—C₁-C₄-alkyl, —SO₂—N(C₁-C₄-alkyl)₂,        —CO—C₁-C₄-alkyl, —O—CO—C₁-C₄-alkyl, —CO—O—C₁-C₄-alkyl, —CO-aryl;        —CO—NH₂, —CO—NH—C₁-C₄-alkyl, —CO—N(C₁-C₄-alkyl)₂;    -   A represents C₁-C₄-alkyl, C₃-C₇-cycloalkyl, halogen-substituted        C₃-C₇-cycloalkyl, aryl, heteroaryl, heterocyclyl,        —NH—CO—C₁-C₄-alkyl, —N(C₁-C₄-alkyl)—CO—C₁-C₄-alkyl, —NH—CO-aryl,        —N(C₁-C₄-alkyl)—CO-aryl, —N(C₁-C₄-alkyl)—CO—C₃-C₇-cycloalkyl,        —NH—CO—NH₂, —NH—CO—NH—C₁-C₄-alkyl, —NH—CS—NH—C₁-C₄-alkyl,        —NH—CO—NH-aryl, —NH—CS—NH-aryl, —SO₂—C₁-C₄-alkyl, —SO₂-aryl, or        —SO₂-heteroaryl; wherein aryl, heteroaryl and heterocyclyl are        optionally substituted with one or more substituents        independently selected from the group consisting of C₁-C₄-alkyl,        C₁-C₄-alkoxy, C₁-C₄-thioalkoxy, ethylenedioxy, CN, halogen or        phenyl, said phenyl being optionally substituted with one or        more substituents independently selected from C₁-C₄-alkyl,        C₁-C₄-alkoxy and halogen atom; and    -   n represents an integer from 0 to 4, inclusive; and    -   and pharmaceutically acceptable salts thereof.

One group of compounds of the invention comprises those compoundswherein W represents COOH.

Another group of compounds of the invention comprises those compoundswherein Y represents NH.

Another group of compounds of the invention comprises those compoundswherein Y represents O.

Another group of compounds of the invention comprises those compoundswherein Y represents N—C₁-C₄-alkyl, especially N—CH₃.

Still another group of compounds of the invention comprises thosecompounds wherein Z represents O.

Still another group of compounds of the invention comprises thosecompounds wherein Z represents S.

Still another group of compounds of the invention comprises thosecompounds wherein Z represents N—C₁-C₄-alkyl, especially N—CH₃.

Still another group of compounds of the invention comprises thosecompounds wherein Z represents N-phenyl.

Still another group of compounds of the invention comprises thosecompounds wherein X represents O.

Still another group of compounds of the invention comprises thosecompounds wherein X represents S.

Still another group of compounds of the invention comprises thosecompounds wherein X represents NSO₂—C₁-C₄-alkyl, especially NSO₂—CH₃.

Still another group of compounds of the invention comprises thosecompounds wherein W represents COOH, Y represents NH, Z represents O andX represents O.

Still another group of compounds of the invention comprises thosecompounds wherein W represents COOH, Y represents O, Z represents O, andX represents O.

Still another group of compounds of the invention comprises thosecompounds wherein W represents COOH, Y represents NH, Z represents O,and X represents NSO₂—C₁-C₄-alkyl, especially NSO₂—CH₃.

Still another group of compounds of the invention comprises thosecompounds wherein W represents COOH, Y represents NH, Z represents S,and X represents NSO₂—C₁-C₄-alkyl, especially NSO₂—CH₃.

A particular embodiment of the compounds of formula (I) as defined aboveare those compounds wherein each of R₁ to R₈ represents hydrogen atom.

Another particular embodiment of the compounds of formula (I) as definedabove are those compounds wherein n is equal to 1 or 2.

Another group of compounds of the invention comprises those compoundswherein A represents. heterocyclyl, said heterocyclyl being optionallysubstituted with one or more substituents independently selected fromthe group consisting of C₁-C₄-alkyl, C₁-C₄-alkoxy, C₁-C₄-thioalkoxy, CN,halogen atom, and phenyl.

Within the above group, A preferably represents isoxazolyl, optionallysubstituted with one or more substituents independently selected fromC₁-C₄-alkyl, especially —CH₃.

Further group of compounds of the invention comprises those compoundswherein A represents phenyl, said phenyl being optionally substituted,especially with ethylenedioxy group.

Further group of compounds of the invention comprises those compoundswherein A represents —N(C₁-C₄-alkyl)—CO—C₃-C₇-cycloalkyl, especially—N(CH₃)—CO-cyclohexyl.

Further group of compounds of the invention comprises those compoundswherein A represents —N(C₁-C₄-alkyl)—CO-heteroaryl wherein heteroaryl isoptionally substituted with one or more substituents independentlyselected from the group consisting of C₁-C₄-alkyl, C₁-C₄-alkoxy,C₁-C₄-thioalkoxy, CN, halogen atom, phenyl, and phenyl optionallysubstituted with one or more substituents independently selected fromthe group consisting of C₁-C₄-alkyl, C₁-C₄-alkoxy, and halogen atom.

A preferred heteroaryl is pyrimidinyl, optionally substituted with oneor more substituents independently selected from the group consisting ofC₁-C₄-alkyl, C₁-C₄-alkoxy, halogen atom, phenyl, and phenyl optionallysubstituted with one or more substituents independently selected fromthe group consisting of C₁-C₄-alkyl, C₁-C₄-alkoxy, and halogen atom.

As examples of specific compounds of the invention, the following can bementioned:

-   1.    (2S)-2-(1,3-benzoxazol-2-ylamino)-3-[4-(2,3-dihydro-1,4-benzodioxin-6-ylmethoxy)phenyl]propionic    acid,-   2.    (2S)-2-(1,3-benzoxazol-2-ylamino)-3-[4-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl]propionic    acid,-   3. (2S)-2-(1,3-benzoxazol-2-ylamino)-3-[4-(2-[(cyclohexylcarbonyl)    -(methyl)amino]ethoxy)phenyl]propionic acid,-   4.    (2S)-2-(1,3-benzoxazol-2-ylamino)-3-[4-(2-[5-methyl-2-(3,4,5-trimethoxyphenyl)-1,3-oxazol-4-yl]ethoxy)phenyl]propionic    acid,-   5.    (2S)-2-(1,3-benzoxazol-2-ylamino)-3-[4-(4-{2-[[6-(2-chlorophenyl-5-cyano-2-    (methylthio)pyrimidin-4-yl](methyl)amino]ethoxy})phenyl]-propionic    acid,-   6.    (2S)-2-(1,3-benzoxazol-2-ylamino)-3-[4-(2-(2-tert-butyl-5-methyl-1,3-oxazol-4-yl)ethoxy)phenyl]propionic    acid,-   7.    (2S)-2-(1,3-benzoxazol-2-ylamino)-3-[4-(2-(2-tert-butyl-5-methyl-1,3-oxazol-4-yl)ethoxy)phenyl]propionic    acid,-   8.    (2S)-2-(1,3-benzoxazol-2-ylamino)-3-[4-(2-[(cyclohexylcarbonyl)-(methyl)amino]thioethoxy)phenyl]propionic    acid,-   9. (2S)-2-(1,3-benzoxazol-2-ylamino)-3-[4-(2-[(cyclohexylcarbonyl)    -(methyl)amino]ethylmethanesulfonylamino)phenyl]propionic acid, and-   10.    (2S)-2-(1,3-benzoxazol-2-yloxy)-3-[4-(2-[(cyclohexylcarbonyl)(methyl)    -amino]ethoxy)phenyl]propionic acid,    and pharmaceutically acceptable salts thereof.

The compounds of the invention have high affinity to the peroxisomeproliferator-activated receptor gamma (PPARγ). Thus the compoundsdemonstrate the ability to bind to the PPARγ and to modulate itsactivity.

The invention relates also to a pharmaceutical composition comprising atleast one compound of formula (I) as defined above or a,pharmaceutically acceptable salt thereof, optionally in combination withother pharmacologically active ingredient, together with one or morepharmaceutically acceptable carriers and/or excipients.

The invention relates also to a compound of formula (I) as defined abovefor use as a medicament.

The invention further relates to the use of a compound of formula (I) asdefined above or a pharmaceutically acceptable salt thereof for thepreparation of a medicament for the treatment and/or prophylaxis ofdiseases and conditions mediated by peroxisome proliferator-activatedreceptor gamma (PPARγ).

The invention further relates to a method of treatment and/orprophylaxis of diseases and conditions mediated by peroxysomeproliferator-activated receptor gamma (PPARγ) in a mammal subject inneed thereof, said method comprising administration to said mammal acompound of formula (I) as defined above in a therapeutically orprophylactically effective amount.

Such PPARγ-mediated diseases and conditions include in particularimpaired insulin tolerance, insulin resistance, type 1 and type 2diabetes, complications resulting from or associated with diabetes, suchas peripheral neuropathy, renal insufficiency, retinopathy,dyslipidemia, syndrome X associated disorders, such as hypertension,obesity, hyperglycemia, atherosclerosis, myocardial ischemia, coronaryheart disease, and other cardiovascular diseases, and renal diseases.

The compounds of the invention can be also useful for improvingcognitive functions.

DETAILED DISCLOSURE OF THE INVENTION

Definitions

The term ,,bioisoster” as used herein relates to a chemical moiety,which replaces another moiety in a molecule of an active compoundwithout significant influence on its biological activity. Otherproperties of the active compound, such as for example its stability asa medicament, can be affected in this way.

As bioisoster moieties for carboxy (COOH) group can be mentionedespecially 5-membered heterocyclic groups having from 1 to 4 heteroatomsselected from nitrogen, oxygen and sulphur, such as for example1,3,4-oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,5-oxadiazolyl,1,2,4-oxadiazolyl, 1,3,4-thiadiazolyl, 1,2,4-thiadiazolyl,1,2,3-thiadiazolyl, 1,2,5-thiadiazolyl, furyl, thienyl, pyrrolyl,pyrazolyl, imidazolyl, isoxazolyl, isothiazolyl, and N-substitutedtetrazolyl. 5-Membered heterocyclic groups can be optionally substitutedwith 1 or 2 substituents selected from the group comprising phenyl,pyridinyl, straight or branched alkyl group, amino group, hydroxy group,fluoro, chloro, bromo, iodo, trifluoromethyl, trifluoromethoxy,trifluorothiomethoxy, alkoxy, and thioalkoxy.

As bioisoster moieties for carboxy (COOH) group can be also mentionedphenyl and 6-membered heterocyclic groups having from 1 to 4 heteroatomsselected from nitrogen, oxygen and sulphur, such as for example pyridyl,pyridinyl, pyrazinyl, pyridazinyl, pyrimidinyl, triazinyl, tetrazinyl,and others. Phenyl and 6-membered heterocyclic groups can be optionallysubstituted with 1 or 2 substituents selected from the group comprisingphenyl, pyridinyl, straight or branched alkyl group, amino group,hydroxy group, fluoro, chloro, bromo, iodo, trifluoromethyl,trifluoromethoxy, trifluorothiomethoxy, alkoxy, and thioalkoxy.

The term “halogen” relates to an atom selected from F, Cl, Br and I.

The term “alkyl” relates to a saturated, straight or branchedhydrocarbon group, having indicated number of carbon atoms. As specificalkyl substituents, the following can be mentioned: methyl, ethyl,propyl, 1-methylethyl, butyl, 1-methylpropyl, 2-methylpropyl,1,1-dimethylethyl, pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl,1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl,1-ethylpropyl, hexyl, 1-methylpentyl, 2-methylpentyl, 1-ethylbutyl,2-ethylbutyl, 3,3-dimethylbutyl, heptyl, 1-ethylpentyl, octyl, nonyl,and decyl.

The term “aryl” relates to a mono- or bicyclic aromatic group, havingfrom 6 to 14 carbon atoms. The examples of aryl groups are phenyl,tolyl, xylyl, naphthyl, such as naphth-1-yl, naphth-2-yl,1,2,3,4-tetrahydronaphth-5-yl, and 1,2,3,4-tetrahydronaphth-6-yl.

The term “heteroaryl” relates to a mono- or bicyclic heteroaromaticgroup, having from 5 to 13 carbon atoms and 1 to 4 heteroatoms selectedfrom N, O, and S. The examples of heteroaryl groups are pyrrol-1-yl,pyrrol-2-yl, pyrrol-3-yl, furyl, thienyl, imidazolyl, oxazolyl,thiazolyl, isoxazolyl, 1,2,4-triazolyl, oxadiazolyl, thiadiazolyl,tetrazolyl, pyridinyl, pyrimidinyl, 1,3,5-triazinyl, indolyl,benzo[b]furyl, benzo[b]thienyl, indazolyl, benzimidazolyl, azaindolyl,cynnolyl, isoquinolinyl, and carbazolyl.

The term “cycloalkyl” relates to a saturated or partially unsaturatedcyclic hydrocarbon group, having from 3 to 7 carbon atoms. The examplesof cycloalkyl groups are cyclopropyl, cyclobutyl, cyclopentyl,cyclopentenyl, cyclopentadienyl, cyclohexyl, cyclohexenyl, andcycloheptyl.

The term “heterocyclyl” relates to a saturated or partially unsaturated5- to 6-membered cyclic hydrocarbon group, having from 1 to 4heteroatoms, selected from N, O and S. Preferred saturated or partiallyunsaturated cyclic hydrocarbon is monocyclic and includes 4 or 5 carbonatoms and 1 to 3 heteroatoms. The examples of heterocyclyl groups arepiperidinyl, piperazinyl, morpholinyl, and pyrrolidinyl.

The compounds of the invention possess chiral center at the carbon atombearing W group and can exist in the form of the respective enantiomers,enantiomer mixtures as well as racemic mixtures.

Therefore, the R and S enantiomers, enantiomer mixtures as well asracemic mixtures of the compounds of formula (I) form the part of theinvention.

Thus in one specific embodiment, the invention relates to compounds offormula (I) having the stereochemical configuration such as shown informula (IA):

wherein W, X, Y, Z, A, n, and R₁ to R₈ have the same meanings as definedabove for formula (I), and pharmaceutically acceptable salts thereof.

In the second specific embodiment, the invention relates to compounds offormula (I) having the stereochemical configuration such as shown informula (IB):

wherein W, X, Y, Z, A, n, and R₁ to R₈ have the same meanings as definedabove for formula (I), and pharmaceutically acceptable salts thereof.

The compounds of formula (I), bearing a basic group, can be convertedinto salts with inorganic or organic acids in a conventional and knownmanner, by the treatment with suitable acid in organic solvent, such asalcohol, ketone, ether or chlorinated solvent, and the recovery of asalt in a conventional manner. Examples of such salts are those withpharmaceutically acceptable inorganic or organic acids. As examples ofinorganic acid salts hydrochloride, hydrobromide, nitrate, sulfate,hydrogensulfate, pyrosulfate, sulfite, pyrosulfite, phosphate,monohydrogenphosphate, dihydrogenphosphate, metaphosphate, andpyrophosphate, can be mentioned. As examples of organic acid saltsacetate, propionate, acrylate, 4-hydroxybutyrate, caprylate, capronate,decanoate, oxalate, malonate, succinate, glutarate, adipate, pimelate,maleate, fumarate, citrate, tartrate, lactate, phenylacetate, mandelate,sebacate, suberate, benzoate, phthalate, alkyl- and arylsulfonates, suchas methanesulfonate, propanesulfonate, p-toluenesulfonate,xylenesulfonate, salicylate, cinnamate, glutamate, aspartate,glucuronate, and galacturonate can be mentioned.

The compounds of formula (I) bearing an acidic group can be convertedinto salts with inorganic or organic base in a conventional and knownmanner by the reaction of a compound of formula (I) with suitableorganic or inorganic base. Salts with pharmaceutically acceptable basesinclude alkaline or alkaline earth metal salts, such as Li, Na, K, Mg orCa, ammonium salts, and salts with basic organic compounds, such as forexample arginine, histidine, piperidine, morpholine, piperazine,ethylenediamine or triethylamine, as well as quaternary ammonium salts.

The present invention relates also to pharmaceutical compositions,comprising a compound of formula (I) with pharmaceutical excipients,depending on the selected route of administration.

One of the embodiments of the invention are pharmaceutical compositionssuitable for oral administration. Pharmaceutical compositions suitablefor oral administration can be in the form of tablets, capsules, pills,lozenges, powders or granules, or solutions or dispersions in a liquid,or similar. Each of said forms will comprise a predetermined amount of acompound of the invention as an active ingredient. The composition inthe form of a tablet can be prepared employing any pharmaceuticalexcipients known in the art for that purpose, and conventionally usedfor the preparation of solid pharmaceutical compositions. The examplesof such excipients are starch, lactose, microcrystalline cellulose,magnesium stearate and binders, for example polyvinylpyrrolidone.Furthermore, an active compound can be formulated as controlled-releasepreparation, such as tablets comprising hydrophilic or hydrophobicmatrix.

Pharmaceutical composition in the form of a capsule can be formulatedusing conventional procedures, for example by incorporation of a mixtureof an active compound and excipients into hard gelatin capsules.Alternatively, a

where X, Y, Z, A, n, and R₁ to R₈ have the meanings as defined forformula (I) above, and W has the meaning other than —COOH or—COO—C₁-C₄-alkyl.

Said substitution can be performed by means of Mitsunobu reaction of thecompound of formula (II) as defined above with a compound of formulaA(CH₂)_(n)—OH wherein A and n have the meanings as defined above,according to the scheme 1:

Mitsunobu reaction can be carried out in anhydrous solvents such asether or halogenated alkane, in the presence of diazo compounds such asDEAD, DIAD, ADDP, and triphenylphosphine, typically at −20 to 20° C.

Alternatively, said substitution of hydrogen atom at X can be performedby alkylating a compound of formula (II) wherein X, Y, Z, and R₁ to R₈have the meanings as defined for formula (I) above, and W has themeaning other than —COOH or —COO—C₁-C₄-alkyl, with a compound of formulaA(CH₂)_(n)—V wherein A and n have the meanings as defined above forformula (I), and V represents a leaving group selected from halogens andalkylsulfonyl or arylsulfonyl groups, in the presence of a strong basecapable of generating the anion from the compound (II), such as forexample sodium hydride, to form a compound of formula (I), according tothe scheme 2:

Alkylation reaction can be performed in an inert organic solvent, suchas anhydrous DMF, THF, DMSO. The strong base capable of generating theanion can be sodium hydride. Sodium hydride can be used dry or as asuspension in mineral oil. Generating of the anion is carried out atroom temperature until the completion of the evolution of hydrogen. Thenin the second stage the alkylating agent A(CH₂)_(n)—V is added, neat oras a solution in an inert organic solvent such as DMF, THF, DMSO. Thesecond step of alkylation can be carried out at 0 to 100° C.

The compounds of the invention of formula (I) wherein W represents —COOHor —COO—C₁-C₄-alkyl, and X, Y, Z, A, n, and R₁ to R₈ have the meaningsas defined above for formula (I), can be prepared by:

a) a substitution of hydrogen atom at X with A(CH₂)_(n)— group in acompound of formula (III)

wherein R represents C₁-C₄ alkyl group, and X, Y, Z, and R₁ to R₈ havethe meanings as defined for formula (I) above, to form a compound offormula (I) wherein W represents an ester group —COOR wherein Rrepresents a C₁-C₄ alkyl group, and X, Y, Z, A, n, and R₁ to R₈ have themeanings as defined for formula (I) above, followed by

b) optionally, a basic hydrolysis of the ester group —COOR to —COOHgroup, to form a compound of formula (I) wherein W represents —COOH.

Said substitution in step a) can be performed by Mitsunobu reaction of acompound of formula (III) with a compound of formula A(CH₂)_(n)—OHwherein A and n have the meanings as defined above for formula (I), toform a compound of formula (IV), according to the scheme 3:

Mitsunobu reaction can be carried out as described above, in anhydroussolvents such as ether or halogenated alkane, in the presence of diazocompounds such as DEAD, DIAD, ADDP, and triphenylphosphine, usually at−20 to 20° C.

Alternatively, said substitution of hydrogen atom at X can be carriedout by reacting a compound of formula (III) wherein R represents C₁-C₄alkyl, and X, Y, Z, and R₁ to R₈ have the meanings as defined forformula (I) above, with a compound of formula A(CH₂)_(n)—V whereinA(CH₂)_(n)— has the meaning as defined above for formula (I), and Vrepresents a leaving group selected from halogens and alkylsulfonyl orarylsulfonyl groups, in the presence of a strong base capable ofgenerating an anion from the compound (III), such as sodium hydride, toform a compound of formula (IV), according to the scheme 4:

The reaction can be carried out as described above for the preparationof compounds of formula (I) wherein W has the meanings other than COOHor —COO—C₁-C₄-alkyl.

The hydrolysis of the ester group in step b) can be carried out in basicconditions, in the manner known in the art. As the examples of the base,alkaline metal hydroxides can be mentioned, such as sodium, potassiumand Lithium hydroxides. For preparing single enantiomers of a compoundof formula (I), it is preferable to carry out the hydrolysis withlithium hydroxide, which allows for the retention of the configuration.

Basic hydrolysis in step b) can be for example carried out in athree-solvent system consisting of THF (tetrahydrofuran), methanol andwater, which allows to obtain homogenous reaction mixture. At the end ofthe hydrolysis, the reaction mixture can be neutralized withhydrochloric acid and, if desired, the free acid product can beextracted, for example with ethyl acetate, according to the scheme 5shown below:

Compounds of formula (I) wherein Y=S, and X, W, Z, A, n, and R₁ to R₈have the meanings as defined above, can be prepared by reaction of acompound of formula (V) wherein W, X, A, n, and R₁ to R₄ have themeanings as defined above for formula (I), with a compound of formula(VI) wherein Z and R₅ to R₈ have the meanings as defined above forformula (I), in the presence of a base in an alcoholic solution,according to the scheme 6.

In the case of the preparation of compounds of formula (I) wherein Wrepresents COOH group, the starting compound in the above process is acompound of formula (V) wherein W is an ester-protected COOH group, asillustrated in the scheme 7. At the end of the reaction, COOH group isdeprotected by basic hydrolysis.

According to the scheme 7, the first reaction step, giving ethyl2-chloro-3-phenylpropionate derivatives, is performed according to themethod described by Y. Kawamatsu, H. Asakawa, T. Saraie, E. Imamiya, K.Nishikawa, Y. Hamuro, Arzneim.Forsch./Drug Res./, 30 (I), 4, 1980,585-589. Chloroester obtained in the Meerwein reaction is reacted with1,3-benzoxazole-2-thiol, in the presence of a base in an alcoholicsolution, to give corresponding ethyl α-(1,3-benzoxazol-2-ylthio)ester.This ester is hydrolyzed in the NaOH or KOH aqueous-alcoholic solution.Free acids are released from salts with diluted hydrochloric acid.

In an analogous manner, the following exemplary compounds were obtained.

Compounds of formula (I) can be prepared both in a racemic form and in aform of a single enantiomer, when starting from optically activematerials. Alternatively, racemic compounds of formula (I) can beresolved into enantiomers, using conventional techniques known in theart.

Tyrosine derivatives of formula (III) wherein X=O, Y=NH, and Z=O, wereobtained according to Shyam B. Advani, Joseph Sam, Journal ofPharmaceutical Sciences, Vol. 57, 10, 1968. For example, according tothe scheme 8, L-tyrosine methyl ester hydrochloride was obtained byesterification of L-tyrosine with methanol in the presence of thionylchloride, followed by the reaction of L-tyrosine methyl esterhydrochloride with 2-chloro-1,3-benzoisoxazole in benzene in thepresence of triethylamine. Similar procedures were used in the case ofD-tyrosine and D,L-tyrosine.

Tyrosine compounds of formula (III) wherein X=O, Y=NH, and Z=NH,N-alkyl, N-aryl, N-heteroaryl or S, can be prepared by adapting themethod of Shyam B. Advani, Joseph Sam, Journal of PharmaceuticalSciences, Vol. 57, 10, 1968, described above.

Tyrosine derivatives of formula (III) wherein X=O, Y=NH, and Z=S, can beprepared according to the method described in Edward S. Lazer, Clara K.Miao, Hin-Chor Wong, Rondla Sorcek, Denice M. Spero, Alex Galman, KollolPal, Mark Behnke, Anne G. Graham, Jane M. Watrous, Carol A. Homon,Juergen Nagle, Arvind Shah, Yvan Guindon, Peter R. Farina, Julian Adams,J. Med. Chem., 1994, 37, 913-923, according to the scheme 9.

4-Mercaptophenytalanine derivatives of formula (III) wherein Y=NH, Z=O,and X=S, were prepared according to the scheme 10, from4-mercaptophenylalanine, which was obtained according to Helen S. M. Lu,Martin Volk, Yuriy Kholodenko, Edward Gooding, Robin M. Hochstrasser,William F. DeGrado, Journal of the American Chemical Society, 119, 31,1997, 7173-7180. The mercapto (SH) group in 4-mercaptophenylalanine wasprotected with trityl group, followed by substitution of one hydrogenatom at α-amino nitrogen atom with 2-benzoxazolyl. The final step of thesynthesis is deprotection of the SH group.

The 4-aminophenylatanine derivatives of formula (III) wherein Y=NH, Z=O,and X=NSO₂—CH₃, were obtained according to the scheme 11 from4-nitro-N-phthaloylphenylalanine methyl ester. The first step of thesynthesis was performed according to F. Bergel, J. A. Stock, Journal ofOrganic Chemistry, 1956, 90-96. 4-Amino-N-phthaloylphenylalanine methylester thus obtained was mesylated with mesyl chloride in pyridine in thepresence of catalytic amounts of DMAP. The subsequent step was theremoval of phthaloyl group, by heating with 6M aqueous HCl. Thusobtained 4-methanesulfonyl-aminophenylalanine was converted into methylester hydrochloride by esterification in methanol in the presence ofthionyl chloride. The subsequent step was the reaction of4-methanesulfonylaminophenylalanine methyl ester hydrochloride with2-chlorobenzoxazole in the presence of triethylamine in benzene.

Starting compounds of formula (VI) wherein Z=O, i.e. substituted2-mercaptobenzoxazoles can be obtained according to Roger Lok, Rondla E.Leone, Antony J. Williams, J. Org. Chem., 61, 3289-3297, in the reactionof a compound of formula (VII) wherein R₅ to R₈ have the meanings asdefined for formula (I) above, as illustrated on the scheme 12.

Starting compounds of formula (VIII), i.e. substituted2-chloro-benzoxazoles can be obtained by using or adapting proceduresdescribed in Fortuna Haviv, James D. Ratajczyk, Robert W. DeNet, FrancisA. Kerdesky, Rolad L. Walters, Steven P. Schmidt, James H. Holmes,Patrick R. Young, George W. Carter, J. Med. Chem., 1988, 31, 1719-1728,by reaction of a compound of formula (VI) wherein R₅ to R₈ have themeanings as defined for formula (I) above, with phosphorus pentoxide,according to the scheme 13.

3-[4-(Benzyloxy)phenyl]-2-hydroxypropionic acid ethyl ester was obtainedaccording to Takamura Makoto, Yanagisawa Hiroaki, Kanai Motoru,Shibasaki Masakatsu, Efficient Synthesis of Antihyperglycemic(S)-α-Aryloxy-β-phenylpropionic Amides Using a Bifunctional AsymmetricCatalyst, Chem. Pharm. Bull., 50, 8, 2002, 1118-1121. Subsequently, theester was treated with sodium hydride and then with 2-chlorobenzoxazole,according to the scheme 14.

The following abbreviations are used herein:

-   -   DIAD: diisopropyl azodicarboxylate    -   DEAD: diethyl azodicarboxylate    -   ADDP: azodicarbonyldipiperidine

EXAMPLES Example 1(2S)-2-(1,3-Benzoxazol-2-ylamino)-3-[4-(2,3-dihydro-1,4-benzodioxin-6-yl-methoxy)phenyl]propionicacid and its methyl ester

R₁ to R₈=H, W=COOH/COOCH₃, X=O, Z=O, Y=NH, n=1,A=2,3-dihydro-1,4-benzodioxin-6-yl of the formula:

2,3-Dihydro-1,4-benzodioxin-6-ylmethanol (0.25 g, 1.5 mmol), methyl(2S)-2-(1,3-benzoxazol-2-ylamino)-3-(4-hydroxyphenyl)propionate (0.31 g,1 mmol) and triphenylphosphine (0.26 g, 1 mmol) were dissolved in 5 mlof tetrahydrofuran (THF). The reaction mixture was cooled to 5° C. DIAD(0.61 g, 3 mmol) was then added and the reaction was stirred at roomtemperature for 18-24 h. Subsequently, THF was evaporated to obtain theproduct, the title acid methyl ester.

The crude product was dissolved in a THF/MeOH/H₂O mixture (6:0.1:1, 2ml). 1M LiOH (1.6 ml) was added and the reaction was stirred for 3 daysat room temperature. Then the reaction mixture was neutralized with 1MHCl, a small amount of water was added and the mixture was extractedwith ethyl acetate. The solvent was evaporated. The purification wasperformed by chromatography. The yield was 50%. MS (ES) 446 (M⁺, 100%)

Example 2(2S)-2-(1,3-Benzoxazol-2-ylamino)-3-[4-((3,5-dimethylisoxazol-4-yl)methoxy)-phenyl]propionicacid and its methyl ester

R₁ to R₈=H, W=COOH/COOCH₃, Y=NH, X=O, Z=O, n=1,A=3,5-dimethylisoxazol-4-yl of the formula:

(3,5-Dimethylisoxazol-4-yl)methanol (0.28 g, 1.5 mmol), methyl(2S)-2-(1,3-benzoxazol-2-ylamino)-3-(4-hydroxyphenyl)propionate (0.31 g,1 mmol) and triphenylphosphine (0.26 g, 1 mmol) were dissolved in 5 mlof tetrahydrofuran (THF). The reaction mixture was cooled to 5° C. DEAD(0.52 g, 3 mmol) was then added and the reaction was stirred at roomtemperature for 18-24 h. Subsequently, THF was evaporated to obtain theproduct, the title acid methyl ester.

The crude product was dissolved in a THF/MeOH/H₂O mixture (6:0.1:1, 2ml). 1M LiOH (1.6 ml) was added and the reaction was stirred for 3 daysat room temperature. Then the reaction mixture was neutralized with 1MHCl, a small amount of water was added and the mixture extracted withethyl acetate. The solvent was evaporated. The purification wasperformed by chromatography. The yield was 60%. MS (ES) 407 (M⁺, 100%)

Example 3(2S)-2-(1,3-Benzoxazol-2-ylamino)-3-[4-(2-[(cyclohexylcarbonyl)(methyl)-amino]ethoxy)phenyl]propionicacid and its methyl ester

R₁ to R₈=H, W=COOH/COOCH₃, X=O, Z=O, Y=NH, n=2,A=(cyclohexyl-carbonyl)methylamino group of the formula:

N-(2-Hydroxyethyl)-N-methylocyclohexanecarboxyamide (0.19 g, 1.5 mmol),methyl (2S)-2-(1,3-benzoxazol-2-ylamino)-3-(4-hydroxyphenyl)-propionate(0.31 g, 1 mmol) and triphenylphosphine (0.26 g, 1 mmol) were dissolvedin 5 ml of tetrahydrofuran (THF). The reaction mixture was cooled to 5°C. ADDP (0.76 g, 3 mmol) was then added and the reaction was stirred atroom temperature for 18-24 h. Subsequently, THF was evaporated to obtainthe product, the title acid methyl ester.

The crude product was dissolved in a THF/MeOH/H₂O mixture (6:0.1:1; 2ml). Aqueous 1M LiOH solution (1.6 ml) was added and the reaction wasstirred for 3 days at room temperature. Then the reaction mixture wasneutralized with 1M HCl, a small amount of water was added and themixture extracted with ethyl acetate. The solvent was evaporated. Thepurification was performed by chromatography. The yield was 40%. MS (ES)465 (M⁺, 100%)

Example 4(2S)-2-(1,3-Benzoxazol-2-ylamino)-3-[4-(2-[5-methyl-2-(3,4,5-trimethoxy-phenyl)-1,3-oxazol-4-yl]ethoxy)phenyl]propionicacid and its methyl ester

R₁ to R₈=H, W=COOH/COOCH₃, X=O, Z=O, Y=NH, n=2,A=[5-methyl-2-(3,4,5-trimethoxyphenyl)-1,3-oxazol-4-yl] of the formula:

To the solution of2-[4-methyl-2-(3,4,5-trimethoxyphenyl)-1,3-oxazol-5-yl]ethanol (2.93 g,10 mmol) in 30 ml of pyridine 4-toluenesulfonyl chloride (1.9 g, 10mmol) was added portionwise at room temperature. Subsequently, thereaction mixture was stirred for 5 h at room temperature, and thenpoured into 200 ml of water and extracted (3×) with 50 ml ofdichloromethane. The combined extracts were washed with 1M HCl, anaqueous solution of sodium bicarbonate, and brine. The organic phase wasdried over magnesium sulfate and the solvent was evaporated, to obtain aproduct, 2-[4-methyl-2-(3,4,5-trimethoxyphenyl)-1,3-oxazol-5-yl]ethyl4-toluenesulfonate having the purity of ca. 95%.

To the solution of 3.12 g of methyl(2S)-2-(1,3-benzoxazol-2-ylamino)-3-(4-hydroxyphenyl)propionate in 50 mlof dimethylformamide 60% suspension of NaH in mineral oil (0,4 g) wasadded portionwise with stirring at room temperature under argonatmosphere. When the evolution of the gas ceased, the solution of2-[4-methyl-2-(3,4,5-trimethoxyphenyl)-1,3-oxazol-5-yl]ethyl4-toluenesulfonate (4.47 g, 10 mmol) in dimethylformamide was addeddropwise. The mixture was heated at 80° C. with stirring. After cooling,the mixture was poured into 1 l of water and extracted several timeswith ethyl acetate. The combined extracts were washed with brine, driedover magnesium sulfate and the solvent was evaporated, to obtain crudemethyl(2S)-2-(1,3-benzoxazol-2-ylamino)-3-[4-(2-[5-methyl-2-(3,4,5-trimethoxy-phenyl)-1,3-oxazol-4-yl]ethoxy)phenyl]propionate.

2.9 g of the crude reaction product obtained above were dissolved in aTHF/MeOH/H₂O mixture (6:0.1:1, 20 ml). 1M LiOH (8 ml) was added and thereaction mixture was stirred for 3 days at room temperature. Then thereaction mixture was neutralized with 1M HCl, a small amount of waterwas added and the mixture extracted with ethyl acetate. The solvent wasevaporated. The purification was performed by chromatography. The yieldwas 40%. MS (ES) 573 (M⁺, 100%)

Example 5(2S)-2-(1,3-Benzoxazol-2-ylamino)-3-[4-(4-{2-[[6-(2-chlorophenyl)-5-cyano-2-(methylthio)pyrimidin-4-yl](methyl)amino]ethoxy})phenyl]propionicacid and its methyl ester

R₁ to R₈=H, W=COOH/COOCH₃, X=O, Z=O, Y=NH, n=2,A=[6-(2-chlorophenyl)-5-cyano-2-(methylthio)pyrimidin-4-yl](methyl)aminogroup of the formula:

4-(2-Chlorophenyl)-6-[(2-hydroxyethyl)(methyl)amino]-2-(methylthio)-pyrimidine-5-carbonitrile(0.50 g, 1.5 mmol), methyl(2S)-2-(1,3-benzoxazol-2-ylamino)-3-(4-hydroxyphenyl)propionate (0.31 g,1 mmol) and triphenyl-phosphine (0.26 g, 1 mmol) were dissolved in 5 mlof tetrahydrofuran (THF). The reaction mixture was cooled to 5° C. DEAD(0.52 g, 3 mmol) was then added and the reaction was stirred at roomtemperature for 18-24 h. Subsequently, THF was evaporated to obtain theproduct, the title acid methyl ester.

The crude product was dissolved in a THF/MeOH/H₂O mixture (6:0,1:1, 2ml). 1M LiOH (1.6 ml) was added and the reaction was stirred at roomtemperature for 3 days. Then the reaction mixture was neutralized with1M HCl, a small amount of water was added and the mixture extracted withethyl acetate. The solvent was evaporated. The purification wasperformed by chromatography. The yield was 58%. MS (ES) 614 (M⁺, 100%)

Example 6(2S)-2-(1,3-Benzoxazol-2-ylamino)-3-[4-(2-(2-tert-butyl-5-methyl-1,3-oxazol-4-yl)ethoxy)phenyl]propionicacid and its methyl ester

R₁ to R₈=H, W=COOH/COOCH₃, X=O, Z=O, Y=NH, n=2,A=2-tert-butyl-5-methyl-1,3-oxazol-4-yl of the formula:

2-(2-tert-Butyl-4-methyl-1,3-oxazol-5-yl)ethanol (0,27 g, 1,5 mmol),methyl (2S)-2-(1,3-benzoxazol-2-ylamino)-3-(4-hydroxyphenyl)propionate(0.31 g, 1 mmol) and triphenylphosphine (0.26 g, 1 mmol) were dissolvedin 5 ml of tetrahydrofuran (THF). The reaction mixture was cooled to 5°C. DEAD (0.52 g, 3 mmol) was then added and the reaction was stirred atroom temperature for 18-24 h. Subsequently, THF was evaporated to obtainthe product, the title acid methyl ester.

The crude product was dissolved in a THF/MeOH/H₂O mixture (6:0.1:1, 2ml). 1M LiOH (1.6 ml) was added and the reaction was stirred at roomtemperature for 3 days. Then the reaction mixture was neutralized with1M HCl, a small amount of water was added and the mixture extracted withethyl acetate. The solvent was evaporated. The purification wasperformed by chromatography. The yield was 55%. MS (ES) 463 (M⁺, 100%)

Example 7(2S)-2-(1,3-Benzoxazol-2-ylamino)-3-[4-(2-[(cyclohexylcarbonyl)(methyl)-amino]thioethoxy)phenyl]propionicacid and its methyl ester

R₁ to R₈=H, W=COOH/COOCH₃, X=S, Z=O, Y=NH, n=2,

A=(cyclohexylcarbonyl)(methyl)amino group of the formula:

N-(2-Hydroxyethyl)-N-methylcyclohexanecarboxyamide (0.19 g, 1.5 mmol),methyl (2S)-2-(1,3-benzoxazol-2-ylamino)-3-(4-mercaptophenyl)-propionate(0.33 g, 1 mmol) and triphenylphosphine (0.26 g, 1 mmol) were dissolvedin 5 ml of tetrahydrofuran (THF). The reaction mixture was cooled to 5°C. DEAD (0.52 g, 3 mmol) was then added and the reaction was stirred atroom temperature for 18-24 h. Subsequently, THF was evaporated to obtainthe product, the title acid methyl ester.

The crude product was dissolved in a THF/MeOH/H₂O mixture (6:0.1:1, 2ml). 1M LiOH (1.6 ml) was added and the reaction was stirred at roomtemperature for 3 days. Then the reaction mixture was neutralized with1M HCl, a small amount of water was added and the mixture extracted withethyl acetate. The solvent was evaporated. The purification wasperformed by chromatography. The yield was 46%. MS (ES) 481 (M⁺, 100%)

Example 8(2S)-2-(1,3-benzoxazol-2-ylamino)-3-[4-(2-[(cyclohexylcarbonyl)(methyl)-amino]ethylmetanesulfonylamino)phenyl]propionicacid and its methyl ester

R₁ to R₈=H, W=COOH/COOCH₃, X=CH₃SO₂N, Z=O, Y=NH, n 2,

A=(cyclohexylcarbonyl)(methyl)amino group of the formula:

N-(2-hydroxyethyl)-N-methylcyclohexanecarboxyamide (0.19 g, 1.5 mmol),methyl(2S)-2-(1,3-benzoxazol-2-ylamino)-3-(4-methanesulfonylamino-phenyl)propionate(0.39 g, 1 mmol) and triphenylphosphine (0.26 g, 1 mmol) were dissolvedin 5 ml of tetrahydrofuran (THF). The reaction mixture was cooled to 5°C. DEAD (0.52 g, 3 mmol) was then added and the reaction was stirred atroom temperature for 18-24 h. Subsequently, THF was evaporated to obtainthe product, the title acid methyl ester.

The crude product was dissolved in a THF/MeOH/H₂O mixture (6:0.1:1, 2ml). 1M LiOH (1.6 ml) was added and the reaction was stirred at roomtemperature for 3 days. Then the reaction mixture was neutralized with1M HCl, a small amount of water was added and the mixture extracted withethyl acetate. The solvent was evaporated. The purification wasperformed by chromatography. The yield was 35%. MS (ES) 542 (M⁺, 100%)

Example 9(2S)-2-(1,3-Benzothiazol-2-ylamino)-3-[4-(2-[(cyclohexytcarbonyl)(methyl)-amino]ethoxy)phenyl]propionicacid and its methyl ester

R₁ to R₈=H, W=COOH/COOCH₃, X=O, Z=S, Y=NH, n=2,

A=(cyclohexylcarbonyl)(methyl)amino group of the formula:

N-(2-hydroxyethyl)-N-methytcyclohexanecarboxyamide (0.19 g, 1.5 mmol),methyl(2S)-2-(1,3-benzothiazol-2-ylamino)-3-(4-hydroxyphenyl)-propionate (0.33g, 1 mmol) and triphenylphosphine (0.26 g, 1 mmol) were dissolved in 5ml of tetrahydrofuran (THF). The reaction mixture was cooled to 5° C.DEAD (0.52 g, 3 mmol) was then added and the reaction was stirred atroom temperature for 18-24 h. Subsequently, THF was evaporated to obtainthe product, the title acid methyl ester.

The crude product was dissolved in a THF/MeOH/H₂O mixture (6:0.1:1, 2ml). 1M LiOH (1.6 ml) was added and the reaction was stirred at roomtemperature for 3 days. Then the reaction mixture was neutralized with1M HCl, a small amount of water was added and the mixture extracted withethyl acetate. The solvent was evaporated. The purification wasperformed by chromatography. The yield was 48%. MS (ES) 481 (M⁺, 100%)

Example 10(2S)-2-(1,3-benzoxazol-2-yloxy)-3-[4-(2-[(cyclohexytcarbonyl)(methyl)-amino]ethoxy)phenyl]propionicacid and its methyl ester

R₁ to R₈=H, W=COOH/COOCH₃, X=O, Z=O, Y=O, n=2

A=(cyclohexylcarbonyl)methylaminoethyl of the formula

N-(2-Hydroxyethyl)-N-methylcyclohexanecarboxyamide (0.19 g, 1.5 mmol),ethyl 2-(1,3-benzoxazol-2-yloxy)-3-(4-hydroxyphenyl)propionate (0.33 g,1 mmol) and triphenylphosphine (0.26 g, 1 mmol) were dissolved in 5 mlof tetrahydrofuran (THF). The reaction mixture was cooled to 5° C. DEAD(0.52 g, 3 mmol) was then added and the reaction was stirred at roomtemperature for 18-24 h. Subsequently, THF was evaporated to obtain theproduct, the title acid methyl ester.

The crude product was dissolved in a THF/MeOH/H₂O mixture (6:0.1:1, 2ml). 1M LiOH (1.6 ml) was added and the mixture was at room temperaturefor 3 days. Then the reaction mixture was neutralized with 1M HCl, asmall amount of water was added and the mixture extracted with ethylacetate. The solvent was evaporated. The purification was performed bychromatography. The yield was 40%. MS (ES) 466 (M+, 100%)

BIOLOGICAL TESTS

The ability of the compounds of the invention to bind to the PPAR gammareceptor and to modify its activity was determined using the followingmethods.

In vitro binding

The ability of the compounds to bind to the PPAR gamma receptor (invitro) was determined according to the procedure described below, usingthe method of competitive radioligand displacement from theligand-receptor complex. PPAR agonist ³H-rosiglitazone at finalconcentration 10 nM was used as the radioligand. An excess of unlabelledtest compounds at final concentration 20 μM was also added to thereaction. The source of the receptor in assays was human recombinantprotein containing LBD (ligand binding domain) of the PPAR gamma. Theseparation of the radioligand unbound with the receptor was performed bydextran coated charcoal technique. The radioactivity was measured usingLS 6500-Beckman Coulter scintillation counter. The obtainedscintillation counts values were compared to the values obtained forsamples incubated with the radioligand (assumed 0% displacement) and tothe values obtained for samples containing both the radioligand and anexcess of non-radiolabelled rosiglitazone (assumed 100% displacement).The obtained values were comprised in the 0-130% range.

References:

-   1. ADD1 /SREBP1 activates PPAR gamma through the production of    endogenous ligand. Proc. Natl. Acad. Sci. USA. 1998 Apr.    14;95(8):4333-7.-   2. An antidiabetic thiazolidinedione is a high affinity ligand for    peroxisome proliferator-activated receptor gamma (PPAR gamma). J.    Biol. Chem. 1995 Jun. 2;270(22): 12953-6.-   3. Fatty acids and eicosanoids regulate gene expression through    direct interactions with peroxisome proliferator-activated receptors    alpha and gamma. Proc. Natl. Acad. Sci. USA. 1997 Apr. 29;    94(9):4318-23.    Binding in Adipocytes

To confirm the ability of the tested molecules to bind in vivo,analogous experiments with the use of murine fibroblasts 3T3-L1 cellline differentiated into adipocytes were performed. Differentiation offibroblasts cells was performed on 12-well plates during 10 days period.On the day of the experiment, the cells were washed twice with PBSsolution prior to 1 h incubation in DMEM medium containingtritium-labelled reference compound (rosiglitazone) at 3 pMconcentration) and different concentrations of the tested compounds (inthe 100 pM-20 μM concentration range) at 37° C. Then the cells werewashed three times with PBS solution and solubilized in 1M NaOHsolution. In the lysate prepared as described above, both radioactivity(using LS 6500 Beckman Coulter scintillation counter) and proteinconcentration (using Bradford method) were measured. Nonspecific bindingwas estimated in the presence of non-labelled reference compound (at 20μM concentration).

The obtained scintillation counts values were compared to the valuesobtained for samples incubated with the radioligand (assumed 0%displacement) and to the values obtained for samples containing both theradioligand and an excess of non-radiolabelled rosiglitazone (assumed100% displacement). The obtained values were comprised in the 0-130%range.

References:

-   1. Identification of high-affinity binding sites for the insulin    sensitizer rosiglitazone (BRL-49653) in rodent and human adipocytes    using a radioiodinated ligand for peroxisomat proliferator-activated    receptor gamma. J. Pharmacol. Exp. Ther. 1998 February;284(2):751-9.-   2. Differential regulation of the stearoyl-CoA desaturase genes by    thiazolidinediones in 3T3-L1 adipocytes. J. Lipid Res. 2000    August;.41(8):1310-6.-   3. Distinct stages in adipogenesis revealed by retinoid inhibition    of differentiation after induction of PPARgamma. Mol Cell Biol. 1996    April;16(4):1567-75.-   4. Differentiation Kinetics of in vitro 3T3-L1 Preadipocyte    Cultures. Tissue Eng. 2002 December;8(6):1071-1081.-   5. Role of PPARgamma in regulating a cascade expression of    cyclin-dependent kinase inhibitors, p18(INK4c) and p21(Waf1/Cip1),    during adipogenesis. J. Biol. Chem. 1999 Jun. 11;274(24): 17088-97.    Adipogenesis

3T3-L1 cell line cells (from ATCC) were maintained in Dulbecco'sModified Eagle's Medium supplemented with 10% Fetal Bovine Serum andantibiotics. Two days before the experiment, the cells were passagedinto 12-well microplates (30×10⁴ cells/well) and maintained forsubsequent 2 days to confluency. After this time, the medium wasreplaced with DMEM+FBS+antibiotics and tested compounds at finalconcentration of 50 μM were added to the cells. Under these conditions,the cells were maintained for 14 days, changing the medium with the testcompounds every 2 days. After 10-14 days the differentiated cells werestained with Oil Red O prior to photographing. References:

-   1. Differential regulation of the stearoyl-CoA desaturase genes by    thiazolidinediones in 3T3-L1 adipocytes. J. Lipid Res. 2000    August;41(8):1310-6.    Glucose Uptake

Differentiated 3T3-L1 fibroblasts were incubated in DMEM supplementedwith 10% FBS and antibiotics with test compounds (at the concentrationof 2 μM) for 48 h. After this time, the cells were washed with PBS, andthen serum-free DMEM was added to the cells. The cells were kept in anincubator for 3 h (37° C./5% CO₂) and then medium was replaced with KHRbuffer (25 mM HEPES-NaOH; pH 7.4; 125 mM NaCl; 5 mM KCl; 1.2 mM MgSO₄;1.3 mM CaCl₂; 1.3 mM KH₂PO₄) and the cells were incubated for 30 minutesat 37° C. Glucose uptake was initiated by the addition to each test wellof 50 μl KRH buffer containing 0,5 mM 2 deoxy-D-[1,2-³H]glucose (0,5μCi) and 100 nM insulin. After 10 min incubation at 37° C., the mediumwas aspirated, and the cells were washed three times with ice-cold KRHbuffer. Then the cells were dissolved in 1M NaOH. In the lysate preparedas described above, both radioactivity (using LS 6500 Beckman Coulterscintillation counter) and protein concentration (using Bradford method)were measured. Nonspecific binding was estimated in the presence ofnon-labelled reference compound (at 20 μM concentration).

References:

-   1. Role of peroxisome proliferator-activated receptor-gamma in    maintenance of the characteristics of mature 3T3-L1 adipocytes.    Diabetes. 2002 July;51(7):2045-55.-   2. Identification of high-affinity binding sites for the insulin    sensitizer rosiglitazone (BRL-49653) in rodent and human adipocytes    using a radioiodinated ligand for peroxisomat proliferator-activated    receptor gamma. J. Pharmacol. Exp. Ther. 1998 February;    284(2):751-9.-   3. Identification of bioactive molecules by adipogenesis profiling    of organic compounds. J. Biol. Chem. 2003 Feb. 28;278(9):7320-4.    Epub 2002 Dec. 19.-   4. Evidence for the involvement of vicinal sulfhydryl groups in    insulin-activated hexose transport by 3T3-L1 adipocytes. J. Biol.    Chem. 1985 Mar. 10;260(5):2646-52.

1. New 3-phenylpropionic acid derivatives of formula (I):

wherein: W represents COOH group or its bioisosters, or —COO—C₁-C₄-alkylgroup; Y represents NH, N—C₁-C₁₀-alkyl, O, or S; Z represents NH,N—C₁-C₁₀-alkyl, N-aryl, N-heteroaryl, S, or O; X represents O, S, NH,N—C₁-C₁₀-alkyl, N-aryl, NSO₂—C₁-C₁₀-alkyl, N—SO₂-aryl, orN—SO₂-heteroaryl; R₁ to R₈ each independently represents hydrogen atomor a substituent selected from the group consisting of: C₁-C₄-alkyl,C₁-C₄-alkoxy, C₃-C₇-cycloalkyl, C₃-C₇-cycloalkoxy, C₁-C₄-thioalkoxy,C₃-C₇-cyclothioalkoxy, halogen atom, halogen-substitutedC₃-C₇-cycloalkyl, aryl, heteroaryl, —NO₂, —CN, —SO₂—NH₂,—SO₂—NH—C₁C₄-alkyl, —SO₂—N(C₁-C₄-alkyl)₂, —CO—C₁-C₄-alkyl,—O—CO—C₁-C₄-alkyl, —CO—O—C₁-C₄-alkyl, —CO-aryl, —CO—NH₂,—CO—NH—C₁-C₄-alkyl, —CO—N(C₁-C₄-alkyl)₂; A represents C₁-C₄-alkyl,C₃-C₇-cycloalkyl, halogen-substituted C₃-C₇-cycloalkyl, aryl,heteroaryl, heterocyclyl, —NH—CO—C₁-C₄-alkyl,—N(C₁-C₄-alkyl)—CO—C₁-C₄-alkyl, —NH—CO-aryl, —N(C₁-C₄-alkyl)—CO-aryl,—N(C₁-C₄-alkyl)—CO—C₃-C₇-cycloalkyl, —NH—CO—NH₂, —NH—CO—NH—C₁-C₄-alkyl,—NH—CS—NH—C₁-C₄-alkyl, —NH—CO—NH-aryl, —NH—CS—NH-aryl, —SO₂—C₁-C₄-alkyl,—SO₂-aryl, or —SO₂-heteroaryl; wherein aryl, heteroaryl and heterocyclylare optionally substituted with one or more substituents independentlyselected from the group consisting of C₁-C₄-alkyl, C₁-C₄-alkoxy,C₁-C₄-thioalkoxy, ethylene-dioxy, CN, halogen or phenyl, said phenylbeing optionally substituted with one or more substituents independentlyselected from C₁-C₄-alkyl, C₁-C₄-alkoxy and halogen atom; and nrepresents an integer from 0 to 4, inclusive; and pharmaceuticallyacceptable salts thereof.
 2. The compound of claim 1 wherein Wrepresents COOH.
 3. The compound of claim 1 wherein Y represents NH. 4.The compound of claim 1 wherein Y represents O.
 5. The compound of claim1 wherein Y represents N—C₁-C₄-alkyl, especially N—CH₃.
 6. The compoundof claim 1 wherein Z represents O.
 7. The compound of claim 1 wherein Zrepresents S.
 8. The compound of claim 1 wherein Z representsN—C₁-C₄-alkyl, especially N—CH₃.
 9. The compound of claim 1 wherein Zrepresents N-phenyl.
 10. The compound of claim 1 wherein X represents O.11. The compound of claim 1 wherein X represents S.
 12. The compound ofclaim 1 wherein X represents NSO₂—C₁-C₄-alkyl, especially NSO₂-CH₃. 13.The compound of claim 1 wherein W represents COOH, Y represents NH, Zrepresents O, and X represents O.
 14. The compound of claim 1 wherein Wrepresents COOH, Y represents O, Z represents O, and X represents O. 15.The compound of claim 1 wherein W represents COOH, Y represents NH, Zrepresents O, and X represents NSO₂—C₁-C₄-alkyl, especially NSO₂—CH₃.16. The compound of claim 1 wherein W represents COOH, Y represents NH,Z represents S, and X represents NSO₂—C₁-C₄-alkyl, especially NSO₂—CH₃.17. The compound of any one of claims 1 to 16 wherein each of R₁ to R₈represents hydrogen atom.
 18. The compound of any one of claims 1 to 17wherein n is equal to 1 or
 2. 19. The compound of any one of claims 1 to18 wherein A represents heterocyclyl, said heterocyclyl being optionallysubstituted with one or more substituents independently selected fromthe group consisting of C₁-C₄-alkyl, C₁-C₄-alkoxy, C₁-C₄-thioalkoxy, CN,halogen atom, and phenyl.
 20. The compound of claim 19 wherein Arepresents isoxazolyl, optionally substituted with one or moresubstituents independently selected from C₁-C₄-alkyl, especially —CH₃.21. The compound of any one of claims 1 to 18 wherein A representsphenyl, said phenyl being optionally substituted with ethylenedioxygroup.
 22. The compound of any one of claims 1 to 18 wherein Arepresents —N(C₁-C₄-alkyl)—CO—C₃-C₇-cycloalkyl.
 23. The compound ofclaim 22 wherein A represents —N(CH₃)—CO-cyclohexyl.
 24. The compound ofany one of claims 1 to 18 wherein A represents—N(C₁-C₄-alkyl)—CO-heteroaryl, said heteroaryl being optionallysubstituted with one or more substituents independently selected fromthe group consisting of C₁-C₄-alkyl, C₁-C₄-alkoxy, C₁-C₄-thioalkoxy, CN,halogen atom or phenyl, said phenyl being optionally substituted withone or more substituents independently selected from C₁-C₄-alkyl,C₁-C₄-alkoxy, and halogen.
 25. The compound of claim 24 whereinheteroaryl is pyrimidinyl, optionally substituted with one or moresubstituents independently selected from the group consisting ofC₁-C₄-alkyl, C₁-C₄-alkoxy, halogen atom, and phenyl, said phenyl beingoptionally substituted with one or more substituents independentlyselected from C₁-C₄-alkyl, C₁-C₄-alkoxy, and halogen atom.
 26. Thecompound of any one of claims 1 to 25 having the stereochemicalconfiguration as shown in formula (IA):

and pharmaceutically acceptable salts thereof.
 27. The compound of anyone of claims 1 to 25 having the stereochemical configuration as shownin formula (IB):

and pharmaceutically acceptable salts thereof.
 28. The compound of claim1, said compound being selected from the following:(2S)-2-(1,3-benzoxazol-2-ylamino)-3-[4-(2,3-dihydro-1,4-benzodioxin-6-ylmethoxy)phenyl]propionicacid,(2S)-2-(1,3-benzoxazol-2-ylamino)-3-[4-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl]propionicacid,(2S)-2-(1,3-benzoxazol-2-ylamino)-3-[4-(2-[(cyclohexylcarbonyl)-(methyl)amino]ethoxy)phenyl]propionicacid,(2S)-2-(1,3-benzoxazol-2-ylamino)-3-[4-(2-[5-methyl-2-(3,4,5-trimethoxyphenyl)-1,3-oksazol-4-yl]ethoxy)phenyl]propionicacid,(2S)-2-(1,3-benzoxazol-2-ylamino)-3-[4-(4-{2-[[6-(2-chlorophenyl)-5-cyano-2-(methylthio)pyrimidin-4-yl](methyl)amino]ethoxy})-phenyl]propionicacid,(2S)-2-(1,3-benzoxazol-2-ylamino)-3-[4-(2-(2-tert-butyl-5-methyl-1,3-oxazol-4-yl)ethoxy)phenyl]propionicacid,(2S)-2-(1,3-benzoxazol-2-ylamino)-3-[4-(2-(2-tert-butyl-5-methyl-1,3-oksazol-4-yl)ethoxy)phenyl]propionicacid,(2S)-2-(1,3-benzoxazol-2-ylamino)-3-[4-(2-[(cyclohexylcarbonyl)-(methyl)amino]thioethoxy)phenyl]propionicacid,(2S)-2-(1,3-benzoxazol-2-ylamino)-3-[4-(2-[(cyclohexylcarbonyl)-(methyl)amino]ethylmethanesulfonylamino)phenyl]propionicacid, and (2S)-2-(1,3-benzoxazol-2-yloxy)-3-[4-(2-[(cyclohexylcarbonyl)-(methyl)amino]ethoxy)phenyl]propionic acid, and pharmaceuticallyacceptable salts thereof.
 29. New 3-phenylpropionic acid derivatives offormula (I):

wherein: W represents COOH; Y represents NH, N—C₁-C₁₀-alkyl, O, or S; Zrepresents NH, N—C₁-C₁₀-alkyl, N-aryl, N-heteroaryl, S, or O; Xrepresents O, S, NH, N—C₁-C₁₀-alkyl, N-aryl, NSO₂—C₁-C₁₀-alkyl,N—SO₂-aryl, or N—SO₂-heteroaryl; each of R₁ to R₈ represents hydrogenatom; A represents C₁-C₄-alkyl, C₃-C₇-cycloalkyl, halogen-substitutedC₃-C₇-cycloalkyl, aryl, heteroaryl, heterocyclyl, —NH—CO—C₁-C₄-alkyl,—N(C₁-C₄-alkyl)—CO—C₁-C₄-alkyl, —NH—CO-aryl, —N(C₁-C₄-alkyl)—CO-aryl,—N(C₁-C₄-alkyl)-—CO—C₃-C₇-cycloalkyl, —NH—CO—NH₂, —NH—CO—NH—C₁-C₄-alkyl,—NH—CS—NH—C₁-C₄-alkyl, —NH—CO—NH-aryl, —NH—CS—NH-aryl, —SO₂—C₁-C₄-alkyl,—SO₂-aryl, or —SO₂-heteroaryl; wherein aryl, heteroaryl and heterocyclylare optionally substituted with one or more substituents independentlyselected from the group consisting of C₁-C₄-alkyl, C₁-C₄-alkoxy,C₁-C₄-thioalkoxy, ethylene-dioxy, CN, halogen or phenyl, said phenylbeing optionally substituted with one or more substituents independentlyselected from C₁-C₄-alkyl, C₁-C₄-alkoxy and halogen atom; and nrepresents an integer from 0 to 4, inclusive; and pharmaceuticallyacceptable salts thereof.
 30. A pharmaceutical composition comprising acompound as defined in claim 1 or a pharmaceutically acceptable saltthereof together with pharmaceutically acceptable carriers and/orexcipients.
 31. A method of treatment and/or prophylaxis of diseases andconditions mediated by peroxysome proliferator-activated receptor gamma(PPARγ) in a mammal subject in need thereof, said method comprisingadministration to said mammal a compound as defined in claims 1 in atherapeutically or prophylactically effective amount.
 32. The method ofclaim 31 wherein said disease or condition is selected from the groupconsisting of type 1 diabetes, type 2 diabetes, insulin resistance,metabolic syndrome, complications resulting from or associated withdiabetes, cardiovascular disorders, atherosclerosis, obesity, cognitivedisorders, and lipid metabolism disorders.